Composition for Anti-inflammation Using an Extract of Scirpus karuizawensis

Abstract

The present invention discloses an anti-inflammatory composition using a pine cone extract having activity that inhibits NO production in a concentration-dependent manner in mouse macrophage cell lines (RAW 264.7 cells) stimulated with LPS (lipopolysaccharide).

Description

Technology Field

[0001] The present invention relates to pine cone sedges ( Scirpus karuizawensis This relates to an anti-inflammatory composition using an extract. Background Technology

[0002] Inflammation is a localized biological defense response that occurs in response to pathological conditions caused by physical trauma, harmful chemicals, infections by bacteria, fungi, or viruses, or irritants among metabolic products. Inflammation is triggered by various inflammatory mediators produced by damaged tissues and migrating cells. During an inflammatory response, plasma accumulates at the site of inflammation to dilute toxins secreted by bacteria, blood flow increases, and symptoms such as erythema, pain, edema, and fever accompany the reaction. Under normal circumstances, the body neutralizes or eliminates causative factors and regenerates damaged tissues through the inflammatory response to restore normal structure and function; however, if this fails, the condition may progress to disease states such as chronic inflammation.

[0003] With the recent advancements in molecular biology, much research has been conducted on inflammatory responses at the molecular level.

[0004] Although various biochemical phenomena are involved in the inflammatory response, macrophages, in particular, are known to play an important role in the inflammatory response by producing nitric oxide (NO) and inflammatory cytokines such as IL-1β, TNF-α, and IL-6 in response to chemical stimuli (Ito T., et al., Curr Drug Target Inflamm Allergy, 2(3):257-265, 2003).

[0005] Nitric oxide is synthesized from L-arginine through the action of nitric oxide synthase (NOS), and NOS exists in several isoforms. bNOS (brain NOS) present in the brain, nNOS (neuronal NOS) present in the nervous system, and eNOS (endothelial NOS) present in the vascular endothelial system are always expressed at a constant level in the body, and the small amount of nitric oxide (NO) produced by these plays an important role in maintaining homeostasis in the human body by performing various physiological reactions related to blood pressure regulation, neurotransmission, learning, and memory. In contrast, iNOS (induced NOS), whose expression is induced by certain stimuli, overproduces NO, and the nitric oxide overproduced by iNOS reacts with superoxide to form peroxynitrite, which acts as a powerful oxidizing agent and damages cells, thereby participating in various pathological processes including inflammation and cancer (Gupta SC et al., Exp Biol Med., 236:658-671, 2011; Riehemann et al., FEBS Lett., 442:89-94, 1999; Stamler et al., Science, 258:1898-1902, 1992).

[0006] Meanwhile, cyclooxygenase (COX) has hydroperoxidase (HOX) activity along with the function of COX and synthesizes the intermediates PGG2 and PGH2 from arachidonic acid, and produces PGE2, PGF2, PGD2, prostacyclin, and thromboxane A2 (TxA2) from these compounds, and there are two isoforms of COX. While COX-1 is always expressed in most tissues and synthesizes prostaglandins (PGs) necessary for cytoprotective action, COX-2 is rapidly induced to be expressed during an inflammatory response and plays an important role in causing the inflammatory response by generating PGE2, etc. (Weisz A., Biochem. J., 316:209-215, 1996; (Miller MJ et al., Mediators of inflammation, 4:387-396, 1995; Appleton L. et al., Adv. Pharmacol., 35:27-28, 1996).

[0007] Non-steroidal anti-inflammatory drugs (NSAIDs), which are currently widely used as anti-inflammatory agents, are known to cause serious side effects such as gastrointestinal disorders, liver damage, and kidney damage (Rainsford KD., Subcell biochem., 42:3-27, 2007; Guruprasad P. Aithal., Rheumatology., 7:139-150, 2011; Praveen PN Rao et al., Pharmaceuticals., 3:1530-1549, 2010). Therefore, research is actively being conducted to find new drugs with sustained effects from natural substances that have anti-inflammatory activity and few side effects.

[0008] The present invention discloses the anti-inflammatory activity of a pine cone sedge extract. The problem to be solved

[0009] The objective of the present invention is to provide an anti-inflammatory composition using pine cone sedge extract.

[0010] Other objects or specific objects of the present invention will be presented below. means of solving the problem

[0011] The present invention was completed by confirming that the pine cone sedge extract inhibits NO production in a concentration-dependent manner in mouse macrophage cell lines (RAW 264.7 cells) stimulated with LPS (lipopolysaccharide), as confirmed in the following examples and experimental examples.

[0012] Considering the foregoing, the present invention can be understood as an anti-inflammatory composition comprising a pine cone sedge extract as an active ingredient in one aspect.

[0013] In this specification, "pine cone sedge extract" refers to an extract obtained by leaching the leaves, branches, roots, bark, underground parts, or mixtures thereof of the pine cone sedge that are the subject of extraction using water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixture thereof using a solvent, an extract obtained using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract. The extraction method may be any method such as cold maceration, reflux, heating, ultrasonic radiation, or supercritical extraction, taking into account the polarity of the active substance, the degree of extraction, and the degree of preservation. The term "fractionated extract" includes fractions obtained by suspending an extract in a specific solvent and then mixing and settling it with a solvent of different polarity, and fractions obtained by adsorbing the crude extract onto a column filled with silica gel or the like and then using a hydrophobic solvent, a hydrophilic solvent, or a mixture thereof as the mobile phase. Furthermore, the term "extract" includes concentrated liquid extracts or solid extracts from which the extraction solvent has been removed by methods such as freeze-drying, vacuum drying, hot air drying, or spray drying. Preferably, it refers to an extract obtained using water, an alcohol having 1 to 4 carbon atoms, or a mixture thereof as the extraction solvent, and more preferably, an extract obtained using a mixture of water and an alcohol having 1 to 4 carbon atoms as the extraction solvent.

[0014] In addition, the term "active ingredient" in this specification means an ingredient that exhibits the intended activity alone or can exhibit activity together with a carrier that is inactive itself.

[0015] Also in this specification, "anti-inflammatory" means improvement (relief of symptoms), treatment, or inhibition or delay of the onset of inflammatory diseases as defined below.

[0016] In addition, in this specification, the term "inflammatory disease" may be defined as a pathological symptom caused by an inflammatory response characterized by a local or systemic biological defense reaction against external physical or chemical stimuli, or infection by external infectious agents such as bacteria, fungi, viruses, and various allergens, or autoimmunity. Such inflammatory response involves a series of complex physiological reactions, such as the activation of various inflammatory mediators and enzymes related to immune cells (e.g., iNOS, COX-2, etc.), the secretion of inflammatory mediators (e.g., secretion of NO, TNF-α, IL-6, etc.), fluid infiltration, cell migration, and tissue destruction, and manifests externally through symptoms such as erythema, pain, edema, fever, and a decline or loss of specific bodily functions. Since the inflammatory disease may be acute, chronic, ulcerative, allergic, or necrotic, any disease is included in the definition of an inflammatory disease as described above, regardless of whether it is acute, chronic, ulcerative, allergic, or necrotic. Specifically, the above inflammatory diseases may include asthma, allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, pulmonary fibroma, irritable bowel syndrome, inflammatory pain, migraine, headache, back pain, fibromyalgia, myofascial disease, viral infection (e.g., hepatitis C), bacterial infection, fungal infection, burn, wound from surgical or dental operation, prostaglandin E hyperactivity syndrome, atherosclerosis, gout, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, scleritis, uveitis, dermatitis (including atopic dermatitis), eczema, multiple sclerosis, etc.

[0017] The anti-inflammatory composition of the present invention may include the active ingredient in any amount (effective amount) as long as it can exhibit activity to improve inflammatory diseases intended for treatment, depending on the specific use, formulation, etc., and the typical effective amount will be determined within the range of 0.001 weight % to 15 weight % based on the total weight of the composition. Here, "effective amount" refers to the amount of the active ingredient that can produce intended medical and pharmacological effects, such as improvement or treatment of inflammatory diseases, or inhibition / delay of the onset of such pathological symptoms, when the composition of the present invention is administered to mammals, preferably humans, the subjects of application, for a period of administration as recommended by medical professionals, etc. Such effective amount may be determined experimentally within the ordinary capacity of a person skilled in the art.

[0018] In addition to the active ingredient, the composition of the present invention may further include any compound or natural extract known in the art to have the corresponding activity and whose safety has already been verified, in order to enhance or reinforce the anti-inflammatory effect or to improve the convenience of taking or consuming by adding similar activities such as anti-allergic activity or skin protective activity (inhibition of skin damage caused by ultraviolet rays, skin moisturization, etc.).

[0019] These compounds or extracts include compounds or extracts listed in pharmacopoeias of various countries (the "Korean Pharmacopoeia" in Korea) and health functional food codes of various countries (in Korea, the "Standards and Specifications for Health Functional Foods," a notification by the Ministry of Food and Drug Safety); compounds or extracts that have received product approval in accordance with the laws of each country governing the manufacture and sale of pharmaceuticals (the "Pharmaceutical Affairs Act" in Korea); and compounds or extracts whose functionality has been individually recognized in accordance with the laws of each country governing the manufacture and sale of health functional foods (the "Act on Health Functional Foods" in Korea). For example, MSM (dimethylsulfonylmethane) with the functionality of "arthritis improvement" in the Korean Health Functional Food Code, N-acetylglucosamine with both "arthritis improvement" and "skin moisturization" functionalities, and compounds or extracts individually recognized for the functionality of "alleviating hypersensitive immune responses" in accordance with the Korean "Act on Health Functional Foods." Enterococcus faecalis A complex of heat-treated dried powder, guava leaf extract, etc., a complex of kiwi extract, perilla leaf extract, picao pretto powder, etc., PLAG (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol), etc., and individually recognized functional ingredients for 'improvement of hypersensitive skin conditions' L. sakei Probio 65, gamma-linolenic acid-containing oil, fruit and vegetable-derived lactic acid bacteria L.plantarum CJLP133, probiotic ATP, etc. would fall under these compounds or extracts.

[0020] One or more of these compounds or natural extracts may be included together with the active ingredients in the anti-inflammatory composition of the present invention.

[0021] The anti-inflammatory composition of the present invention can be understood as a food composition in specific embodiments. In this case, the use of the present invention can be understood as the inhibition of inflammatory responses.

[0022] The food composition of the present invention can be manufactured in any form, for example, as beverages such as tea, juice, carbonated drinks, and isotonic drinks; processed dairy products such as milk and yogurt; chewing gums; rice cakes, Korean confectionery, bread, cookies, and noodles; and health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and bars.

[0023] In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification, as long as it complies with the regulations in effect at the time of manufacture and distribution. For example, it may be a health functional food under the Korean "Act on Health Functional Foods," or a confectionery, legume, tea, beverage, special dietary food, etc., according to each food type in the Food Code of the Korean "Food Sanitation Act" (Notification of the Ministry of Food and Drug Safety "Standards and Specifications for Food").

[0024] The food composition of the present invention may include food additives in addition to its active ingredients. Food additives can generally be understood as substances added to, mixed with, or permeated into food during the manufacture, processing, or preservation of food; since they are consumed daily and over a long period of time along with food, their safety must be guaranteed. Food additive codes in accordance with national laws governing the manufacture and distribution of food (the "Food Sanitation Act" in Korea) restrictively define food additives with guaranteed safety in terms of composition or function. The Korean Food Additive Code (Notification of the Ministry of Food and Drug Safety, "Standards and Specifications for Food Additives") classifies food additives into chemically synthesized products, natural additives, and mixed preparations in terms of composition, and these food additives are classified into sweeteners, flavorings, preservatives, emulsifiers, acidulants, thickeners, etc., in terms of function.

[0025] Sweeteners are used to impart a suitable sweetness to food, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, natural sweeteners are used, and examples of natural sweeteners include corn syrup solids, honey, sucrose, fructose, lactose, maltose, etc.

[0026] Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, natural ones are used. When natural ones are used, nutritional enhancement can be achieved in addition to flavor. Natural flavoring agents may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, etc., or from green tea leaves, Solomon's seal, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. Additionally, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc., may be used. Natural flavoring agents may be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, etc.

[0027] Calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used as preservatives, and acacia gum, carboxymethylcellulose, xanthan gum, pectin, etc. may be used as emulsifiers, and acetic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. may be used as acidifiers. Acidifiers may be added to the food composition to achieve an appropriate acidity, in addition to for the purpose of enhancing flavor, for the purpose of inhibiting the growth of microorganisms.

[0028] As thickening agents, suspending agents, settling agents, gel-forming agents, puffing agents, etc. may be used.

[0029] In addition to the food additives described above, the food composition of the present invention may include physiologically active substances or minerals known in the art and whose stability as food additives is guaranteed, for the purpose of supplementing and reinforcing functionality and nutritional value.

[0030] Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, etc., and examples of minerals include calcium preparations such as calcium citrate, magnesium preparations such as magnesium stearate, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc.

[0031] The food composition of the present invention may include the food additives described above in an appropriate amount to achieve the purpose of their addition, depending on the product type.

[0032] Regarding other food additives that may be included in the food composition of the present invention, reference may be made to the food codes or food additive codes of each country.

[0033] The composition of the present invention may be understood as a pharmaceutical composition in other specific embodiments.

[0034] The pharmaceutical composition of the present invention may be prepared as an oral or parenteral formulation according to the route of administration by conventional methods known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The route of administration may be any suitable route including a local route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissues, and may also be used in combination of two or more routes. An example of a combination of two or more routes is a case where two or more formulations of drugs according to the route of administration are combined, for example, one drug is administered first via the intravenous route and another drug is administered second via the local route.

[0035] Pharmaceutically acceptable carriers are known in the art according to the route of administration or dosage form, and specifically, one may refer to the pharmacopoeias of each country, including the "Korean Pharmacopoeia."

[0036] When the pharmaceutical composition of the present invention is prepared as an oral formulation, it may be prepared in the form of powder, granules, tablets, pills, coated tablets, capsules, liquids, gels, syrups, suspensions, wafers, etc., in accordance with methods known in the art together with a suitable carrier. Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol; starches such as corn starch, potato starch, and wheat starch; celluloses such as cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinylpyrrolidone; water; methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate; mineral oil; malt; gelatin; talc; polyols; vegetable oils; ethanol; glycerol; etc. Suitable binders, lubricants, disintegrants, coloring agents, diluents, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrite, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweeteners, sodium alginate, polyethylene glycol, wax, etc. Suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium and calcium salts, polyethylene glycol, etc. Suitable disintegrants include starch, methylcellulose, agar, bentonite, xanthan gum, starch, alginic acid, or its sodium salt, etc. Other diluents include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc.

[0037] When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injectable, transdermal, nasal inhalant, and suppository according to methods known in the art with a suitable carrier. When formulated as an injectable, an aqueous isotonic solution or suspension may be used as a suitable carrier; specifically, isotonic solutions such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose may be used. When formulated as a transdermal formulation, it may be formulated in the form of an ointment, cream, lotion, gel, topical solution, paste, liniment, aerosol, etc. In the case of nasal inhalers, they can be formulated in the form of an aerosol spray using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide, and when formulated as suppositories, the carriers may include Witepsol, Tween 61, polyethylene glycols, cocoa starch, laurin starch, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, and sorbitan fatty acid esters.

[0038] Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to literature such as [Remington's Pharmaceutical Sciences (19th ed., 1995)]. The said literature is to be considered part of this specification.

[0039] A preferred dosage of the pharmaceutical composition of the present invention may be in the range of 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g / kg, depending on the patient's condition, body weight, gender, age, severity of the patient, and route of administration. Administration may be administered once a day or divided into several doses. Such dosages shall not be construed as limiting the scope of the present invention in any aspect.

[0040] In another specific embodiment, the composition of the present invention may be understood as a cosmetic composition. If the anti-inflammatory composition of the present invention is understood as a cosmetic composition, its use may be understood as suppressing inflammatory skin troubles, alleviating inflammatory skin irritation, etc.

[0041] Even if the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may take on any product classification in terms of its use or legal classification, and specifically, it may be a functional cosmetic or a non-functional general cosmetic used for purposes such as improving skin troubles or atopic dermatitis. In terms of product form, it may also take on any product form, specifically, it may take the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, spray, etc. In terms of specific product form, it may be a formulation such as a softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.

[0042] The cosmetic composition of the present invention may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, pigments, and fragrances, and carriers.

[0043] In the case where the formulation of the present invention is a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0044] In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0045] When the formulation of the present invention is a solution or an emulsion, a solvent, a solubilizing agent, or an emulsifying agent is used as a carrier component, specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, etc. may be used.

[0046] In the case where the formulation of the present invention is a suspension, liquid diluents such as water, ethanol, or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, etc. may be used as carrier components.

[0047] In the case where the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester, etc. may be used as a carrier component.

[0048] The cosmetic composition of the present invention can be prepared according to the method of preparing cosmetic compositions commonly practiced in the art, except that it includes an active ingredient exhibiting anti-inflammatory activity.

[0049] In another aspect, the present invention relates to an antioxidant composition comprising pine cone sedge extract as an active ingredient.

[0050] With respect to the antioxidant composition of the present invention, the meaning of the extract, effective amount, specific product form, etc., shall be applied exactly as described in the anti-inflammatory composition of the present invention as previously stated. Effects of the invention

[0051] As described above, according to the present invention, an anti-inflammatory composition using pine cone sedge extract can be provided.

[0052] The composition of the present invention can be commercialized into products such as food, cosmetics, and pharmaceuticals for uses such as the improvement of inflammatory diseases. Brief explanation of the drawing

[0053] Figure 1 shows the results of the cytotoxicity and anti-inflammatory activity evaluation (NO assay). Figure 2 is an evaluation of antioxidant activity (DPPH assay). Specific details for implementing the invention

[0054] The present invention will be explained below with reference to examples and experimental examples. However, the scope of the present invention is not limited to these examples and experimental examples.

[0055] <Example> Preparation of Pinus Cone Extract

[0056] Pinecone sedge ( Scirpus karuizawensis Makino) Outpost 500 g of dry powder was cold-macerated twice for 48 hours in 5 L of 50% ethanol. The extract was filtered using a filter (Advantech No. 1), and the filtrate was concentrated under reduced pressure at 40°C to obtain an ethanol extract. The extract concentrated under reduced pressure was freeze-dried to obtain a powder-form extract.

[0057] <Experimental Example> Anti-inflammatory activity experiment and antioxidant activity experiment

[0058] <Experimental Example 1> Anti-inflammatory Activity Experiment - NO assay

[0059] The Raw 264.7 mouse macrophages used in the experiment were purchased from the ATCC (American Type Culture Collection, Manassas, VA, USA). Raw 264.7 mouse macrophages were placed in a 96-well plate at a density of 3 × 10⁶ 4 After dispensing the cells into each well, they were cultured for 24 hours under conditions of 37°C and 5% CO2.

[0060] Subsequently, ethanol extracts of *Sorbus aucuparia* were applied to each culture medium at different concentrations. In the wells excluding the non-mitogenic group, Lipopolysachatide, which acts as an endotoxin and is used as a NO synthesis mitogen, was applied at a concentration of 1 μg / ml for 1 hour to induce NO synthesis. After culturing for 24 hours, the degree of NO secretion inhibition was confirmed. Nitric oxide (NO) was measured by mixing the culture supernatant with the corresponding quantitative reagent in a 1:1 weight ratio and measuring the NO secretion inhibitory effect using 540 nm absorbance.

[0061] The result is the 50% inhibition concentration of NO production (IC10). 50 The results are shown in Table 1 and Figure 1 below. The results are based on three repeated experiments for each concentration.

[0062] Sample IC 50 Pinecone extract 53.31 ㎍ / ml L-NMMA (positive control) 20.20 uM

[0063] As confirmed in the table above, the IC of the pine cone sedge extract 50 It was found to be 53.31 μg / ml, and the IC50 of the positive control, L-NMMA... 50 It was found to be 20.20 uM.

[0064] Although not presented in the experimental results, the cytotoxicity evaluation results according to the MTT assay showed that the pine cone sedge extract did not exhibit any particular cytotoxicity up to 100 μg / ml; therefore, the NO assay evaluation was performed with a maximum treatment concentration of 100 μg / ml (Fig. 1).

[0065] <Experimental Example 2> Antioxidant Activity Experiment - DPPH Method

[0066] After preparing a 200 µM DPPH methanol solution, 100 µL of the DPPH solution and 100 µL of the sample diluted in methanol at each concentration were added and reacted at room temperature for 30 minutes. Subsequently, the absorbance was measured at 517 nm using an ELISA reader, and the DPPH radical scavenging activity of the sample at each treatment concentration was determined by comparing it to the untreated control group according to the formula below, and the concentration corresponding to 50% DPPH scavenging activity (IC10) was calculated. 50 The results are shown in Table 2 and Figure 2 below. BHA, an antioxidant, was used as a positive control. The results are based on three repeated experiments for each concentration.

[0067] Antioxidant activity according to the DPPH method (IC 50 , μg / ml) division DPPH assay (µg / ml) Pinecone extract 47.00 BHA (positive control) 22.37

[0068] As confirmed in the table above, the IC of the pine cone sedge extract 50 It was found to be 47.00 µg / ml, and the IC50 of BHA, the positive control, was 50 It was found to be 22.37 μg / ml.

Claims

Claim 1 An anti-inflammatory composition comprising pine cone sedge extract as an active ingredient. Claim 2 A composition according to claim 1, characterized in that the extract is a water and ethanol mixed solvent extract of the whole plant of *Sorbus aucuparia*. Claim 3 A composition according to claim 1 or 2, characterized in that the composition is a pharmaceutical composition. Claim 4 A composition according to claim 1 or 2, characterized in that the composition is a food composition. Claim 5 A composition characterized in that, in claim 1 or 2, the composition is a cosmetic composition. Claim 6 An antioxidant composition comprising, as an active ingredient, a water, ethanol, or mixed solvent extract of the whole plant of *Sorbus aucuparia*.

Images